PHYSICO-CHEMICAL ANALYSIS AND IMPACT OF DIFFERENT SHODHANA METHODS ON GUGGULU

Objective: Guggulu is one of the important Dravya (drug) used in Ayurvedic formulations since ancient time which means “Protection against diseases”. Ashuddhaguggulu has physical and chemical impurities which need to be eliminated before using in formulation by Shodhana. The present study was conducted to evaluate and compare the effect of different Shodhana methods i.e., Dolayantra Shodhana and dissolving Shodhana on properties of Guggulu by employing various physic-chemical and chromatographic methods. Method: Physicochemical screening was done by evaluating ash, Acid Insoluble Ash, Loss on Drying, Water Soluble Extractive and Alcohol Soluble Extractive. Chromatographic analysis was performed to estimate guggulsterone (E and Z) content, using High Performance Liquid Chromatography (HPLC). Result: The Physico-chemical studies showed decrease in LOD, Ash and Acid Insoluble Ash content and increase in extractive values such as Alcohol Soluble Extractive of Guggulu after Shodhana process. The HPTLC analysis showed significant change in guggulsterone (E and Z) content in Guggulu after Shodhana process Conclusion: This study helps to understand the effect of different Shodhana methods on the efficacy of drug. In this study, we established qualitative profile of Shodhit Guggulu in terms of physicochemical parameters and phytochemical content by HPLC

Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test

Triple null mutations in starch synthase SSIIa gene homoeologs lead to high amylose and resistant starch in hexaploid wheat

Lack of nutritionally appropriate foods is one of the leading causes of obesity in the US and worldwide. Wheat (Triticum aestivum) provides 20% of the calories consumed daily across the globe. The nutrients in the wheat grain come primarily from the starch composed of amylose and amylopectin. Resistant starch content, which is known to have significant human health benefits, can be increased by modifying starch synthesis pathways. Starch synthase enzyme SSIIa, also known as starch granule protein isoform-1 (SGP-1), is integral to the biosynthesis of the branched and readily digestible glucose polymer amylopectin. The goal of this work was to develop a triple null mutant genotype for SSIIa locus in the elite hard red winter wheat variety ‘Jagger’ and evaluate the effect of the knock-out mutations on resistant starch content in grains with respect to wild type. Knock-out mutations in SSIIa in the three genomes of wheat variety ‘Jagger’ were identified using TILLING. Subsequently, these loss-of function mutations on A, B, and D genomes were combined by crossing to generate a triple knockout mutant genotype Jag-ssiia-∆ABD. The Jag-ssiia-∆ABD had an amylose content of 35.70% compared to 31.15% in Jagger, leading to ~ 118% increase in resistant starch in the Jag-ssiia-∆ABD genotype of Jagger wheat. The single individual genome mutations also had various effects on starch composition. Our full null Jag-ssiia-∆ABD mutant showed a significant increase in RS without the shriveled grain phenotype seen in other ssiia knockouts in elite wheat cultivars. Moreover, this study shows the potential for developing nutritionally improved foods in a non-GM approach. Since all the mutants have been developed in an elite wheat cultivar, their adoption in production and supply will be feasible in future.https://doi.org/10.1186/s12870-020-02822-

Mapping of Quantitative Trait Loci for Grain Iron and Zinc Concentration in Diploid A Genome Wheat

Micronutrients, especially iron (Fe) and zinc (Zn), are deficient in the diets of people in underdeveloped countries. Biofortification of food crops is the best approach for alleviating the micronutrient deficiencies. Identification of germplasm with high grain Fe and Zn and understanding the genetic basis of their accumulation are the prerequisites for manipulation of these micronutrients. Some wild relatives of wheat were found to have higher grain Fe and Zn concentrations compared with the cultivated bread wheat germplasm. One accession of Triticum boeoticum (pau5088) that had relatively higher grain Fe and Zn was crossed with Triticum monococcum (pau14087), and a recombinant inbred line (RIL) population generated from this cross was grown at 2 locations over 2 years. The grains of the RIL population were evaluated for Fe and Zn concentration using atomic absorption spectrophotometer. The grain Fe and Zn concentrations in the RIL population ranged from 17.8 to 69.7 and 19.9 to 64.2 mg/kg, respectively. A linkage map available for the population was used for mapping quantitative trait loci (QTL) for grain Fe and Zn accumulation. The QTL analysis led to identification of 2 QTL for grain Fe on chromosomes 2A and 7A and 1 QTL for grain Zn on chromosome 7A. The grain Fe QTL were mapped in marker interval Xwmc382-Xbarc124 and Xgwm473-Xbarc29, respectively, each explaining 12.6% and 11.7% of the total phenotypic variation and were designated as QFe.pau-2A and QFe.pau-7A. The QTL for grain Zn, which mapped in marker interval Xcfd31-Xcfa2049, was designated as QZn.pau-7A and explained 18.8% of the total phenotypic variatio

A diploid wheat TILLING resource for wheat functional genomics

Citation: Rawat, N., . . . & Gill, B. (2012). A diploid wheat TILLING resource for wheat functional genomics. BMC Plant Biology, 12(1), 205. https://doi.org/10.1186/1471-2229-12-205Background: Triticum monococcum L., an A genome diploid einkorn wheat, was the first domesticated crop. As a diploid, it is attractive genetic model for the study of gene structure and function of wheat-specific traits. Diploid wheat is currently not amenable to reverse genetics approaches such as insertion mutagenesis and post-transcriptional gene silencing strategies. However, TILLING offers a powerful functional genetics approach for wheat gene analysis. Results: We developed a TILLING population of 1,532 M[subscript 2] families using EMS as a mutagen. A total of 67 mutants were obtained for the four genes studied. Waxy gene mutation frequencies are known to be 1/17.6 - 34.4 kb DNA in polyploid wheat TILLING populations. The T. monococcum diploid wheat TILLING population had a mutation frequency of 1/90 kb for the same gene. Lignin biosynthesis pathway genes- COMT1, HCT2, and 4CL1 had mutation frequencies of 1/86 kb, 1/92 kb and 1/100 kb, respectively. The overall mutation frequency of the diploid wheat TILLING population was 1/92 kb. Conclusion: The mutation frequency of a diploid wheat TILLING population was found to be higher than that reported for other diploid grasses. The rate, however, is lower than tetraploid and hexaploid wheat TILLING populations because of the higher tolerance of polyploids to mutations. Unlike polyploid wheat, most mutants in diploid wheat have a phenotype amenable to forward and reverse genetic analysis and establish diploid wheat as an attractive model to study gene function in wheat. We estimate that a TILLING population of 5, 520 will be needed to get a non-sense mutation for every wheat gene of interest with 95% probability

Using empirical science education in schools to improve climate change literacy

Providing children with a clear understanding of climate change drivers and their mitigation is crucial for their roles as future earth stewards. To achieve this, it will be necessary to reverse the declining interest in STEM (Science, Technology, Engineering and Mathematics) education in schools in the UK and other countries, as STEM skills will be critical when designing effective mitigation solutions for climate change. The ‘Heat-Cool Initiative’ was co-designed and successfully implemented in five primary/secondary UK schools, as a playful learning tool to unleash student interest in STEM subjects. 103 students from two cohorts (years 5–6 and 7–9) participated in five Heat-Cool activity sessions where they used infrared cameras to explore the issue of urban heat. Their learning was evaluated using a multi-functional quantitative assessment, including pre- and postsession quizzes. Climate change literacy increased by 9.4% in primary school children and by 4.5% in secondary school children. Analyses of >2000 infrared images taken by students, categorised into 13 common themes, revealed age-related differences in children’s cognitive development. At primary school age, images of the ‘self’ dominated; secondary school children engaged more with their physical environment. This novel approach demonstrated the importance of developing tailored technology-enhanced STEM education programmes for different age cohorts, leading to a high capacity for improving learning outcomes regarding climate change. Such programmes, embedded in school curricula nationally and internationally, could become a much-needed positive contribution to reaching the United Nation’s Sustainable Development Goals, especially Goals 4 (Quality Education) and 13 (Climate Action)

TILL-D: An Aegilops tauschii TILLING Resource for Wheat Improvement

Aegilops tauschii (2n = 2x = 14, genome DD), also known as Tausch’s goatgrass, is the D genome donor of bread or hexaploid wheat Triticum aestivum (2n = 2x = 42, AABBDD genome). It is a rich reservoir of useful genes for biotic and abiotic stress tolerance for wheat improvement. We developed a TILLING (Targeting Induced Local Lesions In Genomes) resource for Ae. tauschii for discovery and validation of useful genes in the D genome of wheat. The population, referred to as TILL-D, was developed with ethyl methanesulfonate (EMS) mutagen. The survival rate in M1 generation was 73%, out of which 22% plants were sterile. In the M2 generation 25% of the planted seeds showed phenotypic mutations such as albinos, chlorinas, no germination, variegated, sterile and partially fertile events, and 2,656 produced fertile M2 plants. The waxy gene was used to calculate the mutation frequency (1/70 kb) of the developed population, which was found to be higher than known mutation frequencies for diploid plants (1/89–1/1000 kb), but lower than that for a polyploid species (1/24–1/51 kb). The TILL-D resource, together with the newly published Ae. tauschii reference genome sequence, will facilitate gene discoveries and validations of agronomically important traits and their eventual fine transfer in bread wheat

Integration of genetic and genomics resources in einkorn wheat enables precision mapping of important traits

Einkorn wheat (Triticum monococcum) is an ancient grain crop and a close relative of the diploid progenitor (T. urartu) of polyploid wheat. It is the only diploid wheat species having both domesticated and wild forms and therefore provides an excellent system to identify domestication genes and genes for traits of interest to utilize in wheat improvement. Here, we leverage genomic advancements for einkorn wheat using an einkorn reference genome assembly combined with skim-sequencing of a large genetic population of 812 recombinant inbred lines (RILs) developed from a cross between a wild and a domesticated T. monococcum accession. We identify 15,919 crossover breakpoints delimited to a median and average interval of 114 Kbp and 219 Kbp, respectively. This high-resolution mapping resource enables us to perform fine-scale mapping of one qualitative (red coleoptile) and one quantitative (spikelet number per spike) trait, resulting in the identification of small physical intervals (400 Kb to 700 Kb) with a limited number of candidate genes. Furthermore, an important domestication locus for brittle rachis is also identified on chromosome 7A. This resource presents an exciting route to perform trait discovery in diploid wheat for agronomically important traits and their further deployment in einkorn as well as tetraploid pasta wheat and hexaploid bread wheat cultivars

An overview of monitoring methods for assessing the performance of nature-based solutions against natural hazards

To bring to fruition the capability of nature-based solutions (NBS) in mitigating hydro-meteorological risks (HMRs) and facilitate their widespread uptake require a consolidated knowledge-base related to their monitoring methods, efficiency, functioning and the ecosystem services they provide. We attempt to fill this knowledge gap by reviewing and compiling the existing scientific literature on methods, including ground-based measurements (e.g. gauging stations, wireless sensor network) and remote sensing observations (e.g. from topographic LiDAR, multispectral and radar sensors) that have been used and/or can be relevant to monitor the performance of NBS against five HMRs: floods, droughts, heatwaves, landslides, and storm surges and coastal erosion. These can allow the mapping of the risks and impacts of the specific hydro-meteorological events. We found that the selection and application of monitoring methods mostly rely on the particular NBS being monitored, resource availability (e.g. time, budget, space) and type of HMRs. No standalone method currently exists that can allow monitoring the performance of NBS in its broadest view. However, equipments, tools and technologies developed for other purposes, such as for ground-based measurements and atmospheric observations, can be applied to accurately monitor the performance of NBS to mitigate HMRs. We also focused on the capabilities of passive and active remote sensing, pointing out their associated opportunities and difficulties for NBS monitoring application. We conclude that the advancement in airborne and satellite-based remote sensing technology has signified a leap in the systematic monitoring of NBS performance, as well as provided a robust way for the spatial and temporal comparison of NBS intervention versus its absence. This improved performance measurement can support the evaluation of existing uncertainty and scepticism in selecting NBS over the artificially built concrete structures or grey approaches by addressing the questions of performance precariousness. Remote sensing technical developments, however, take time to shift toward a state of operational readiness for monitoring the progress of NBS in place (e.g. green NBS growth rate, their changes and effectiveness through time). More research is required to develop a holistic approach, which could routinely and continually monitor the performance of NBS over a large scale of intervention. This performance evaluation could increase the ecological and socio-economic benefits of NBS, and also create high levels of their acceptance and confidence by overcoming potential scepticism of NBS implementations

Nature-based solutions efficiency evaluation against natural hazards: modelling methods, advantages and limitations

Nature-based solutions (NBS) for hydro-meteorological risks (HMRs) reduction and management are becoming increasingly popular, but challenges such as the lack of well-recognised standard methodologies to evaluate their performance and upscale their implementation remain. We systematically evaluate the current state-of-the art on the models and tools that are utilised for the optimum allocation, design and efficiency evaluation of NBS for five HMRs (flooding, droughts, heatwaves, landslides, and storm surges and coastal erosion). We found that methods to assess the complex issue of NBS efficiency and cost-benefits analysis are still in the development stage and they have only been implemented through the methodologies developed for other purposes such as fluid dynamics models in micro and catchment scale contexts. Of the reviewed numerical models and tools MIKE-SHE, SWMM (for floods), ParFlow-TREES, ACRU, SIMGRO (for droughts), WRF, ENVI-met (for heatwaves), FUNWAVE-TVD, BROOK90 (for landslides), TELEMAC and ADCIRC (for storm surges) are more flexible to evaluate the performance and effectiveness of specific NBS such as wetlands, ponds, trees, parks, grass, green roof/walls, tree roots, vegetations, coral reefs, mangroves, sea grasses, oyster reefs, sea salt marshes, sandy beaches and dunes. We conclude that the models and tools that are capable of assessing the multiple benefits, particularly the performance and cost-effectiveness of NBS for HMR reduction and management are not readily available. Thus, our synthesis of modelling methods can facilitate their selection that can maximise opportunities and refute the current political hesitation of NBS deployment compared with grey solutions for HMR management but also for the provision of a wide range of social and economic co-benefits. However, there is still a need for bespoke modelling tools that can holistically assess the various components of NBS from an HMR reduction and management perspective. Such tools can facilitate impact assessment modelling under different NBS scenarios to build a solid evidence base for upscaling and replicating the implementation of NBS